Details of new avian and pandemic flu
research projects announced today

IP/06/1413
announces €28.3 million of new funding for research projects in the field
of avian and pandemic influenza. This MEMO outlines the 17 projects selected for
funding. Final budget figures (EU contribution) and project details are subject
to the final signature of contracts, and so may change. A glossary of terms used
in the project descriptions is found at the end of the document.

The primary aim of this project is to
develop better avian influenza vaccines through live or vector vaccines that
could be mass applicable through spray, drinking water or eye drop. These vector
vaccines would offer considerable advantages: mass applicable, less labour
intensive and animal friendly application, protection by local and systemic
immunity and less interference with eventual maternal antibodies, more complete
protection through cellular and humoral immunity, faster onset of immunity when
used in face of an outbreak and cheaper production methods. The project exploits
recently acquired knowledge concerning the molecular characterization of the
viruses resulting in the construction of candidate strains with highly
interesting efficacy and safety profile. Safety and efficacy with Newcastle
disease (NDV) vectors and infectious laryngotracheitis (ILT) vectors both for H5
and for H7 inserts have already been demonstrated in vivo. Such vaccines would
also have marker aspects which will allow differentiation of vaccinated from
infected animals (DIVA principle). Sensitive, specific and easy to use marker
diagnostic tests that will be compatible with the vaccines will also be
developed.

The project focuses on technology transfer and
training and will: i) Organise technical workshop to facilitate technology
transfer particularly in the field of molecular diagnostic tools for pathogen
detection and differentiation, to reinforce epidemiological analysis for
monitoring and modelling of Avian influenza especially and to respond to
outbreaks with infectious diseases of livestock in general. ii) Provide training
through organization of seminars and short term courses in well qualified
laboratories of a number of member states of European Union and iii)

Organize technical workshops, courses and training in the partner countries
to improve the technical experimental level of the staff and laboratories in
charge of livestock infectious diseases. The consortium includes seven European
and seven developing countries as well as FAO/IAEA, OIE and FAO animal health
programmes.

FLU-LAB-NET - Development and enhancement of laboratory networks for
avian influenza

FLU-LAB-NET provides new opportunities for
enhancement and reinforcement of the Community Reference Laboratory and National
Reference Laboratory network for avian influenza (AI) within the EU. This will
strengthen harmonisation and development of laboratory and diagnostic methods,
coordination of research efforts and sharing of expertise. Rapid responses to
national and global emergencies with data sharing will be key areas of
exploitation, contributing to a European laboratory task force capability for AI
in animal species. Rapid, formal interactive communications will be addressed
through web-based forums. Laboratories involved in influenza research in
domestic mammals will also participate. FLU-LAB-NET will also foster formal
links and coordinate with corresponding human, swine and equine influenza
networks. FLU-LAB-NET provides opportunities for identification and development
of the complementarities of global, multi-disciplinary influenza research
programmes. Strategically important third country and INCO partners are also
included in this network, in order to raise laboratory standards and benefit
from knowledge sharing. This will promote greater trust, understanding and early
access to information that may be of importance to both veterinary and public
health in the EU.

Highly pathogenic avian influenza
viruses (HPAIV) have acquired the unprecedented and alarming capability to
infect humans. The presence of HPAIV in wild birds poses a continuous risk for
poultry and fatal human infections. Due to the lack of sufficient knowledge,
attempts to avoid or contain HPAIV outbreaks have been largely unsuccessful.
Full understanding of the ecology and pathogenesis of HPAIV requires a
multidisciplinary approach determining host-pathogen interactions and the role
played by the host immune response. To this end, the FLUPATH consortium was
established. FLUPATH is composed of 6 National Reference Laboratories for avian
influenza, 5 academic institutions and 2 institutions specialized in animal
science and health. The participants, with expertise in chicken genomics,
micro-array technology, pathology, receptors, innate immunity and chicken
immunology, will use multidisciplinary and complementary approaches to address
key problems and unanswered questions with respect to the ecology and
pathogenesis of avian influenza.

FLUPATH will provide knowledge and tools for new strategies which will be
tailored for the control and management of avian influenza at the European and
International level.

Avian influenza (AI)
represents one of the greatest concerns for public health that has emerged in
recent times. The ongoing Asian H5N1 outbreak is a serious concern for food
security and human health world-wide. Evidence is growing that highly pathogenic
H5N1 is not only spreading by poultry trade but is also carried by wild birds.
Recently, H5N1 infected wild birds, mainly waterfowl, have been detected in
several countries in the European Union. These findings indicate that H5N1 is
becoming more and more endemic in wild birds. The finding of a cat, a stone
marten and raptors that died as a result of H5N1 infection, highlight the
consequences of this development. Questions are being raised about the risk of
contamination of surface water for animals and humans.

At present there is a lack of scientific data on the survival of AI viruses
in fresh and processed poultry products, in feathers and down, and in poultry
manure and litter. Because of the lack of data, a proper assessment of the risk
posed by these materials for spreading or reintroduction of disease is
impossible. The objective of this proposal is to generate data on AI virus
stability and the effect of biological, chemical and physical parameters there
upon. Moreover, it will provide knowledge about virus survival under different
physical conditions. This will enable proper risk assessment of the trade in
treated and fresh poultry commodities and poultry litter.

The primary goal of this project
will be the development and application of technologies to combat avian
influenza (AI) infections. A study will be conducted to establish the
effectiveness of the current EU surveillance and early warning systems for AI
and then to develop blueprints for improvements to these programmes in disease
free periods and during outbreaks. The model will include criteria for
harmonised diagnostic tests for on-farm outbreak investigation. To complement
this study a range of diagnostic tools will be developed, evaluated and
validated alongside the evaluation of a range of commercially available tests.
This will include high throughput techniques for molecular and serological
testing, penside testing and simplified tests for use in laboratories with
limited resources or experience. Efforts will particularly focus on the
validation of tests for use on clinical materials derived from Anseriformes,
other wild birds species and some selected mammalian species.

The present world
wide avian influenza crisis has highlighted the need for comprehensive training
and the transfer of technology to accession and INCO countries with the clear
goal of aiding these countries in combating AI with the most up-to-date
diagnostic and disease management procedures. The FLUTRAIN project aims to at
two levels. It will firstly approach the need for training by providing three
workshops over the duration of the project (3 years) that will call on experts
in the AI field to pass on their valuable expertise in the diagnosis and
management of AI to participants from accession and INCO countries. Training
opportunities will also be provided in partner labs in order to consolidate the
information and practical experience gained during the workshops. In addition a
CD-ROM will also be prepared containing essential information provided during
the workshops. A website will be developed which will enable participants and
the general public to access the training programs and will include on-line
discussion fora between trainees and trainers. The second goal of FLUTRAIN will
be the transfer of technology to accession and INCO countries. This will include
the provision of new, simplified and cost effective diagnostic methods and
reagents. It will also involve the transfer of deliverables, both for
serological and virological diagnosis that have been (or will be) developed in
three European projects namely AVIFLU, Lab-on-site and FLUAID. The consortium is
made up of 19 members which includes 2 SMEs. The participants can be identified
as trainers (12) or recipients (7). Given the rapid evolution of AI on a
worldwide level, remaining recipient countries will be identified.

INN-FLU will increase the
understanding of host susceptibility and disease pathogenesis of hpAIV in
chickens. The project is based on four columns: (i) characterization of the
virulence of an hpH5N1 strain isolated from a Thai eagle in several species,
(ii) determination of host factors influenced by AIV infection during the very
early phase of infection, (iii) research on whether different lines of chicken
breeds show differences in resistance to hpH5N1 infection, and (iv)
investigation of contact points between wild birds and domestic free range
poultry and migratory behaviour of birds based on satellite tracking. The
objective is to better understand disease pathogenesis, virulence determinants,
and host-pathogen interactions and to define the molecular basis for host
specificity in chickens. The major focus with regard to host-pathogen
interactions will be on the innate immune response in birds and the role of
individual influenza virus proteins involved. Experiments will also address the
question whether HA molecules from non-H5 and non-H7 strains can give low
pathogenic viruses the potential to become highly pathogenic. The scientifically
outstanding partners of the consortium know each other well and have already
collaborated successfully in earlier research projects. It is to be expected
that the project will deepen our knowledge of influenza virus infection and
transmission in birds substantially.

NEW-FLUBIRD - Network for Early Warning of Influenza Viruses in Migratory
Birds in Europe

NEW-FLUBIRD will establish a European network of
virologists and ornithologists, data managers, epidemiologists and modellers, in
order to provide “early warning and risk assessment systems” in real
time for the threat posed to animal and human health by avian influenza (AI)
viruses from migratory birds. The network will largely build on, and extend
existing collaborations between AI virologists in Europe and international
ornithological organizations active within and outside Europe including Africa,
Middle East and Eastern Europe. Epidemiological assessments will thus cover the
major flyways of migratory birds over Europe and the areas from which migratory
birds in Europe migrate. Furthermore it will focus on experimental infection of
selected migratory bird species with HPAI virus H5N1 and possibly other relevant
HPAI viruses, to determine pathogenesis and excretion profiles. In turn, the
ornithological studies will construct migratory route maps and set up systematic
sampling from healthy wild migratory birds thus providing insight in volume and
timing of migration as well as key sites of those migratory bird species that
pose the highest risk of transmitting HPAI viruses to poultry in Europe.
Finally, NEW-FLUBIRD will seek integration with global early warning systems
developments like GLEWS of FAO and WHO (Global Early Warning System) and GNAIS
of WCS (Global 3 Network for Avian Influenza Surveillance).

Ducks play a major role in the epidemiology of AI since
wild waterfowl, including ducks, constitute the natural reservoir of all
subtypes of influenza and is considered as the “Trojan horse” of
Asian H5N1 HPAI. The NOVADUCK project brings together both private (2 companies,
1 SME) and public sector stakeholders (European reference laboratories and
agencies on AI) from 4 European countries towards the aim of developing and
evaluating new highly protective and cost-effective avian influenza live
vaccines for ducks, based on live viral vectors and in line with the DIVA
strategy (Differentiating between Infected and Vaccinated Animals). Viral
vectors will be engineered to optimise both their immunogenicity and protective
capacities. Vaccine candidates will be pre-screened for safety and
immunogenicity using newly developed duck-specific immunological tools. The best
vaccine candidates will be evaluated for efficacy in a HPAI challenge-model in
ducks. Serological DIVA test able to detect infection in vaccinated duck flocks
will be generated and the effect of vaccination on genetic and antigenic drift
of H5N1 will be assessed.

The RIVERS project aims 1/ to gather data on the
survival of avian influenza viruses, in particular A(H5N1), in the natural
environment; 2/ to generate scientific knowledge about the survival of avian
influenza viruses, in experimental settings; 3/ to provide figures about the
effect of various food processing methods (drying, salting, using acidic
compounds etc .....) on influenza virus survival; and 4/ to elaborate models
about the survival of avian influenza virus, mainly on A(H5N1), in natural
environments to demonstrate their perpetuation in nature both in biological and
environmental reservoirs. Once gathered, and fully analyzed, it is envisaged
that the data corpus will provide researchers, risk managers and policy makers
with an invaluable source for the prevention and control of avian influenza
A(H5N1) at times of epizootics, and endemic but sustained viral circulation and
at times of post crisis management..

The
ultimate objective of the EUROFLU project is to fill the knowledge gaps
concerning the molecular factors and mechanisms of highly pathogenic avian
influenza virus (HPAIV) transmission and pathogenesis. This objective will be
reached through integration of interdisciplinary experimental research
approaches and bioinformatics analyses. The project has three major aspects. One
is to identify, characterize and validate HPAIV-factors that are involved in the
recognition and targeting of the virus to the cellular host receptor. Since the
latter varies between humans and different animals, it determines which species
the virus attacks. The second objective is to reveal viral and cellular factors
and mechanisms that regulate virus replication within the infected cell and can
therefore determine cell tropism (the way the virus enters only certain kinds of
cells and not others) and – again – host specificity. Focus will be
on virus/host-interactions including viral factors from different HPAIV (H5 and
H7, and H5/H7-reassortants). A third aspect is the use of mouse and chicken
models for in vivo analysis of HPAIV infections of mammals and birds to
monitor HPAIV-transmission and pathogenicity within organisms. Overall, the
project covers many of the major aspects relating to the pathogenicity of avian
influenza virus, and the process of flu infection, which in the medium term may
lead to new treatments.

This project aims to identify and study the
essential viral and host factors that determine the outcome of an influenza
infection – so, unlike vaccine projects, which try to get a solution from
the virus side, FLUINNATE approaches the problem from the host side. Influenza A
virus (FLUAV) enters the human respiratory tract and must replicate in the face
of multiple innate immune defence mechanisms to establish infection in
vivo. Successful viruses must adapt and evolve the capacity of circumventing
the body's antiviral interferon (IFN) response. The project will test the
hypothesis that the speed and efficiency by which a given virus circumvents
these early host responses determine its host range and the severity of the
disease it causes. Virus-induced inflammatory mediators (like IFN) exert
powerful effects against FLUAV in the lung. However, they may also worsen the
patient's symptoms. The researchers will analyze viral factors governing the
innate antiviral cytokine response and determine the impact of these factors on
virus growth, cell survival and pathogenicity. Human, avian and pig FLUAV will
be used in animal models and in cell culture systems, such as human airway
lining cells. The present studies should generate important information that
will help to better understand the processes involved in the emergence of lethal
influenza viruses and to develop efficient control measures against these
devastating pathogens.

Currently circulating H5N1 avian influenza viruses
could cause a devastating pandemic if they became transmissible between humans.
It is therefore crucial to understand the mechanisms whereby influenza virus
adapts from avian to human hosts. Several recent studies have highlighted the
importance for transmissibility of mutations in the proteins of the viral
replicative machinery. This project will undertake a comprehensive study of the
molecular structure and function of the influenza virus polymerase with the aim
of understanding how it adapts during inter-species transmission. The
researchers will focus on determination of the atomic structure of two important
viral proteins: polymerase and the trimeric complex. Through advanced functional
genomics, studies in animals and bioinformatic methods they will identify
mutations that are particular important for transmission between different
species. This will help to elaborate new molecular tools for influenza research
and to design strategies for screening novel anti-influenza drugs targeting
polymerase. The overall goal of the project is to provide new knowledge that
will allow to better monitor and combat the emergence of pandemic influenza
strains.

The novel adjuvant IC31TM stimulates both strong T cell
and B cell responses and has a broad mechanism of action as well as an excellent
safety profile. Pre-clinical animal studies with inter-pandemic (seasonal)
influenza vaccines revealed that IC31TM induces long lasting cellular immune
responses characterized by high levels of IFN-producing T cells and increased
haemagglutination inhibition antibodies – a hallmark of the body's defence
against the virus. At the same time, a more than 10-fold reduction in vaccine
(antigen) dose could be achieved. This is important, since vaccine production
capacity would be an important bottleneck in a potential epidemic. The
consortium plans to perform thorough pre-clinical testing in order to prepare
for clinical studies (Phase 1 and human challenge studies) with a whole
inactivated virus vaccine based on a pandemic influenza virus strain (H5N1)
produced in cell culture and adjuvanted with IC31TM. In contrast to the
currently most widely used procedure of influenza virus production in eggs, the
main benefits of cell culture production are in immediate and more reliable
availability, constant quality of substrate and reduced risk of contamination.
Successful clinical studies would provide safety data for the use of the novel
adjuvant IC31TM and proof-of-concept for improved immunogenicity and dose
sparing. In contrast to the common adjuvant Aluminium hydroxide, the combination
of whole virus and IC31TM is expected to provide broader immunity due to the
induction of T cell responses against conserved epitopes.

This project aims to
characterize genetically engineered H5N1 vaccine candidates that are –
through the deletion of a certain protein (NS1) at the same time unable to
replicate and also exert a self-adjuvant effect: The genetically engineered
virus more strongly stimulates the production of certain substances that help
increase the body's immune response. While traditional vaccines need to be
produced in chicken eggs, the vaccine candidates of this consortium will be
produced in cell culture – an easier and faster way to get large
quantities of vaccine. Given intra-nasally, it is expected to provide both
systemic immunity and local immunity at the site of virus entry (on the mucosa
of the respiratory tract). After pre-clinical assessment in ferrets and macaques
(including the development of novel sensitive immunological assays), human phase
I and IIa clinical trials as well as a human challenge study with attenuated
H5N1 virus will be conducted.

PANFLUVAC – Efficacious vaccine formulation system for prophylactic
control of influenza pandemics

The overall aim of PANFLUVAC is
to construct vaccine delivery systems for intranasal and injectable vaccines.
New H5N1 vaccines are to be based on well-established virosome technology -
proven its worth for vaccines against seasonal influenza – as well as
whole virus vaccines. This will permit direct comparison of these promising
approaches. An intranasal vaccine would offer a very convenient and low-tech way
of delivery, and whole virus vaccines promise to activate more broadly the
different "branches" of the human immune system. Vaccine potency will be
enhanced by a novel adjuvant - an agent that enhances the body's immune reaction
to the vaccine - called ISCOMs. PANFLUVAC aims to generate the first intranasal
H5N1 vaccine within the first 12 months of the project. The PANFLUVAC project is
also designed to facilitate rapid modification of the vaccine in the face of
virus drift. Within the preclinical evaluation, the new vaccines will be tested
for the degree of cross protection they offer against different influenza virus
variants. Overall, PANFLUVAC offers a generic vaccine development system to
provide safe and efficacious vaccines against influenza.

Partners from Switzerland, Italy, United Kingdom, Norway, and Germany.
Project includes 3 SMEs.

Glossary

antiviral interferon response = a general defence mechanism of the
body (involving the secretion of a several substances, among them interferons)
against all kinds of viruses

cell tropism = the specificity of the virus for certain kinds of
target cells which it infects (the influenza virus for example infects the cells
lining the airways and lungs, but will not infect muscle cells).

ecology = pattern of relations between organisms and their
environment

epidemiology = all factors controlling the presence or absence of a
disease

host susceptibility = the receptiveness of the human or animal host
for the disease (in this case for example the fact that humans can only be
infected with certain but not with other viruses).

In vivo = in live animals or humans (as opposed to in
vitro = in the test tube, in cell culture)

pathogenesis = the mechanism by which certain factors cause
disease

polymerase = a virus protein that is essential for its replication

self-adjuvant = an adjuvant is a substance that is added to a vaccine
in order to increase the body's immune response to this vaccine. This will
usually allow achieving efficient protection from the disease with a lower
amount of vaccine. In this special case, the genetically engineered vaccine
virus has properties that by themselves (without the addition of an extra
adjuvant) increase the body's immune response.

Virosome = virosomes represent reconstituted empty influenza
virus envelopes that cannot replicate but contain the important antigens HA
(haemagglutinin) and NA (neuraminidase) that elicit the immune response and can
therefore serve as an efficient vaccine.